JP4549806B2 - Automatic cell culture apparatus using autoclave sterilization and method of using the same - Google Patents

Automatic cell culture apparatus using autoclave sterilization and method of using the same Download PDF

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JP4549806B2
JP4549806B2 JP2004309295A JP2004309295A JP4549806B2 JP 4549806 B2 JP4549806 B2 JP 4549806B2 JP 2004309295 A JP2004309295 A JP 2004309295A JP 2004309295 A JP2004309295 A JP 2004309295A JP 4549806 B2 JP4549806 B2 JP 4549806B2
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cell culture
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operation unit
automatic cell
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JP2006115798A (en
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睦 高木
勝己 中嶋
利尚 土井
隆昌 緒方
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Kawasaki Motors Ltd
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Kawasaki Jukogyo KK
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Priority to US11/257,950 priority patent/US7883887B2/en
Priority to DE602005002213T priority patent/DE602005002213T2/en
Priority to AT05256595T priority patent/ATE371719T1/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/02—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using physical phenomena
    • A61L2/04—Heat
    • A61L2/06—Hot gas
    • A61L2/07—Steam
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/50—Means for positioning or orientating the apparatus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/48—Automatic or computerized control
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/809—Incubators or racks or holders for culture plates or containers

Abstract

An automatic cell cultivation apparatus capable of sterilizing completely, including a cold storage unit and a normal temperature storage unit, is used for cell cultivation of plural subjects, without causing cross contamination. An automatic cell cultivation apparatus comprising an operation unit 11 for operating to cultivate cells of subjects, incubator units 14 for cultivating the cells, a cold storage unit 16 for storing reagents necessary for cultivation, a normal temperature storage unit 17 for storing culture tools, an inlet-outlet unit 15 for taking in and out reagents and culture tools, and a steam supply unit 22 for autoclave sterilization, in which the incubator units 14, the cold storage unit 16, the normal temperature storage unit 17, and the inlet-outlet unit 15 communicate with the operation unit 11, and have sealing doors 18a to 18d leading to the operation unit 11, filters 20a and 20b for cleaning and sterilizing the air stream supplied in the operation unit 11 are provided, and sealing doors 30a and 30b are provided between the filter units and the operation unit 11. By selecting the sealing doors 18a to 18d and opening and closing, sterilizing steam from the steam supply unit 22 is supplied into any of the sterilization required units, operation unit 11, and the incubator units 14, the cold storage unit 16, the normal temperature storage unit 17, and the inlet-outlet unit 15, communicating with the operation unit 11.

Description

本発明は、オートクレーブ滅菌を利用した自動細胞培養装置及びその使用方法に関し、更に詳細には、複数の被検者の細胞培養に使用しても交叉汚染が生じることがなく、しかも保管部を含めて完全に滅菌することが可能な自動細胞培養装置及びその使用方法に関する。   The present invention relates to an automatic cell culture apparatus using autoclave sterilization and a method of using the same, and more particularly, there is no cross contamination even when used for cell culture of a plurality of subjects, and the storage unit is included. The present invention relates to an automatic cell culture apparatus capable of being completely sterilized and a method of using the same.

近年、人体の皮膚、軟骨、骨、血管、臓器等の細胞、組織等を取り出して、対外で培養した後、同一人又は他人の治療に使用するという再生医療が発達し、一部では実用化され始めている。このような再生医療に於いては、人体から採取した少量の細胞が体外で培養されるが、この細胞培養に際しては、培養細胞の汚染を防止することが重要である。特に複数の被検者の細胞培養を同時に行う場合には、培養中に他人の細胞やこれに付着していた感染症の細菌、ウィルス、マイコプラズマ等が混入するという交叉汚染を如何にして防止するかが重要な問題となる。   In recent years, regenerative medicine has been developed in which cells, tissues, etc. of human skin, cartilage, bone, blood vessels, organs, etc. are taken out and cultured outside, and then used for the treatment of the same person or others. Being started. In such regenerative medicine, a small amount of cells collected from the human body are cultured outside the body. In this cell culture, it is important to prevent contamination of the cultured cells. In particular, when cell culture of multiple subjects is performed at the same time, how to prevent cross-contamination of other people's cells and infectious bacteria, viruses, mycoplasma, etc. adhering to the cells during the culture. Is an important issue.

一方、再生医療の実用化には安全で低コストの細胞培養が求められており、インキュベーション、培地交換操作、継代操作等を細胞培養完了まで繰り返して自動で行う自動化装置の必要性が検討されている。このような培養操作の自動化に際しては、上述の交叉汚染の問題を避けるため、現状では、一人の被検者に一台の細胞培養装置を専用で細胞培養完了まで使用する、ダウンフローの気流を使用した不完全な汚染対策の下で培養装置を使用する、等の方法が採用されているが、上記の問題の十分な解決策とはなっていない。   On the other hand, safe and low-cost cell culture is required for the practical application of regenerative medicine, and the necessity of an automated device that automatically repeats incubation, medium exchange operation, subculture operation, etc. until cell culture is completed is studied. ing. When automating such culture operations, in order to avoid the above-mentioned cross-contamination problem, at present, the flow of downflow that uses one cell culture device exclusively for one subject until cell culture is completed is used. Although a method such as using a culture device under the used countermeasure against incomplete contamination is adopted, it is not a sufficient solution to the above problem.

即ち、一人に一台の細胞培養装置を専用で使用する場合、他人の細胞はその装置内には存在しないため、培養中に交叉汚染が起こるという問題はない。しかし、装置の利用効率の点からコストが高くなるという問題がある。また、ダウンフローの気流を使用した培養装置では、他人の細胞が同じ装置内に存在することとなり、その細胞やこれに付着している細菌、ウィルス、マイコプラズマ等が他の被検者用の容器などに付着することを完全に防止することができず、交叉汚染の問題の十分な解決策とはなっていない。   That is, when one cell culture apparatus is used exclusively for one person, there is no problem that cross-contamination occurs during the culture because the cells of other people do not exist in the apparatus. However, there is a problem that the cost increases from the viewpoint of the utilization efficiency of the apparatus. In addition, in a culture device that uses a downflow airflow, other people's cells are present in the same device, and the cells, bacteria, viruses, mycoplasma, etc. adhering to these cells are contained in containers for other subjects. It cannot be completely prevented from adhering to the surface and is not a sufficient solution for the problem of cross contamination.

このような問題を解決するため、交叉汚染を防止しつつ複数の被検者の細胞培養を可能とする装置の開発が行われている（特許文献１）。この自動培養装置ではインキュベータ部を複数備え、培地交換操作、継代操作等の培養操作を行う操作部は共通で使用し、一人の被検者の培養操作が終わる度にオゾン等によるガス滅菌が行われるが、これらの残留ガスが装置内に存在すると、培養すべき細胞が死滅するという問題点がある。また、この自動培養装置では、装置本体内のインキュベータ部や操作部は滅菌することができるが、培養装置に付随した、例えば培養に使用する試薬類や培養器具類を保管する保管部、試薬類や培養器具類の搬入及び搬出を行うための入出庫部等を滅菌することはできない。そのため、保管部や入出庫部を含めて完全に交叉汚染を防ぐことはできない。
国際公開第２００４／０１１５９３号パンフレット（図２） In order to solve such a problem, an apparatus that enables cell culture of a plurality of subjects while preventing cross contamination has been developed (Patent Document 1). This automatic culture device is equipped with multiple incubators, and a common operation unit for performing culture operations such as medium exchange and subculture is used, and gas sterilization with ozone or the like is performed each time a culture operation of one subject is completed. However, if these residual gases are present in the apparatus, there is a problem that cells to be cultured are killed. Further, in this automatic culture apparatus, the incubator section and the operation section in the apparatus main body can be sterilized, but a storage section for storing, for example, reagents used for the culture and culture instruments attached to the culture apparatus, and reagents. In addition, it is not possible to sterilize the loading / unloading section for carrying in and out the culture equipment. Therefore, it is impossible to completely prevent cross-contamination including the storage unit and the loading / unloading unit.
International Publication No. 2004/011593 Pamphlet (Figure 2)

本発明は、上述のような問題点を解決するために為されたものであり、本発明の課題は、複数の被検者の細胞培養に使用しても交叉汚染が生じることがなく、しかも保管部、入出庫部等を含めて完全に滅菌することが可能な自動細胞培養装置を提供することである。   The present invention has been made to solve the above-described problems, and the problem of the present invention is that cross-contamination does not occur even when used for cell culture of a plurality of subjects. It is to provide an automatic cell culture device that can be completely sterilized including a storage unit, a loading / unloading unit, and the like.

上記課題を解決するため、本発明のオートクレーブ滅菌を利用した自動細胞培養装置は、被検者の細胞培養に必要な培養操作を行うための操作部と、細胞を培養するためのインキュベータ部と、培養に必要な試薬類や培養器具類を保管する保管部と、オートクレーブ滅菌を行うための蒸気供給部とを備え、前記インキュベータ部、前記保管部は、それぞれ前記操作部に連通すると共に、前記操作部との間にそれぞれ密閉扉を備えており、該密閉扉を選択して開閉することにより、前記操作部と、該操作部に通ずる前記インキュベータ部及び前記保管部の何れかとに前記蒸気供給部からの滅菌蒸気が供給されることを特徴とする。   In order to solve the above problems, an automatic cell culture apparatus using autoclave sterilization of the present invention includes an operation unit for performing a culture operation necessary for cell culture of a subject, an incubator unit for culturing cells, A storage unit for storing reagents and culture tools necessary for culture, and a steam supply unit for performing autoclave sterilization, wherein the incubator unit and the storage unit communicate with the operation unit, and Each of which is provided with a hermetic door, and by selecting and opening and closing the hermetic door, the steam supply unit is connected to the operation unit and either the incubator unit or the storage unit connected to the operation unit. The sterilization steam from is supplied.

本発明の自動細胞培養装置ではオートクレーブ滅菌が採用されているため、ガス滅菌とは異なり残留ガスの心配がないため、短時間で滅菌を完了することが可能である。また、オートクレーブ滅菌は、通常、病院等の医療機関で使用される滅菌装置で採用されており、滅菌効果が高いため、確実に交叉汚染を防止することができる。   Since autoclave sterilization is employed in the automatic cell culture device of the present invention, unlike gas sterilization, there is no concern about residual gas, and sterilization can be completed in a short time. Autoclave sterilization is usually employed in sterilizers used in medical institutions such as hospitals, and since it has a high sterilization effect, cross contamination can be reliably prevented.

また、操作部に通ずるインキュベータ部及び保管部は、密閉扉を開閉することにより、操作部と連通させることができるため、滅菌が必要なときに操作部と共に滅菌を行うことが可能となっている。
また、前記自動細胞培養装置に於いては、試薬及び培養器具類の出し入れを行うための入出庫部を更に備え、前記入出庫部は前記操作部に連通すると共に、前記操作部との間に密閉扉を備えており、前記密閉扉を開閉することにより前記蒸気供給部からの滅菌蒸気が供給されるように構成される。
これにより、入出庫部は密閉扉を開閉することにより、操作部と連通させることができるため、滅菌が必要な時に操作部と共に滅菌を行うことが可能となっている。 In addition, since the incubator part and the storage part leading to the operation part can be communicated with the operation part by opening and closing the sealed door, it is possible to perform sterilization together with the operation part when sterilization is necessary. .
The automatic cell culture apparatus further includes a loading / unloading unit for loading / unloading reagents and culture instruments, and the loading / unloading unit communicates with the operation unit, and between the operation unit. A sealed door is provided, and sterilization steam from the steam supply unit is supplied by opening and closing the sealed door.
Thereby, since the entering / exiting part can be made to communicate with the operation part by opening and closing the sealing door, it is possible to perform sterilization together with the operation part when sterilization is necessary.

また、上記自動細胞培養装置に於いては、前記操作部に供給される気流の除塵及び除菌を行うフィルタ部と、該フィルタ部及び前記操作部の間に設けられた密閉扉とを更に備え、オートクレーブ滅菌に際して前記フィルタ部を前記操作部から遮断するように構成される。   The automatic cell culture apparatus further includes a filter unit that performs dust removal and sterilization of the airflow supplied to the operation unit, and a sealed door provided between the filter unit and the operation unit. The filter unit is configured to be shut off from the operation unit during autoclave sterilization.

これにより、オートクレーブ滅菌に際してフィルタ部が熱や蒸気により損傷を受けることを防止することができる。   Thereby, it is possible to prevent the filter portion from being damaged by heat or steam during autoclave sterilization.

ここで、前記操作部には、操作ロボットと、遠心分離器と、細胞培養に必要な培養操作機器等を設け、前記操作部は、前記操作ロボット及び前記遠心分離器を配置した機器設置部と、前記培養操作機器を配置した培養操作部とを備えた構成とし、前記機器設置部と前記培養操作部との間に遮断装置を設けてもよい。これにより、前記培養操作部のみを滅菌することが可能となる。   Here, the operation unit is provided with an operation robot, a centrifuge, a culture operation device and the like necessary for cell culture, and the operation unit includes an apparatus installation unit in which the operation robot and the centrifuge are arranged. The culture operation unit may be provided with a culture operation unit, and a blocking device may be provided between the device installation unit and the culture operation unit. Thereby, it becomes possible to sterilize only the said culture operation part.

本発明に於いて、前記細胞培養に必要な培養操作機器には、ターンテーブルと、ピペット装置と、遠心管ハンドリング装置とが含まれ、前記細胞培養に必要な前記培養操作には、培地交換操作と継代操作とが含まれる構成とすることができる。   In the present invention, the culture operation equipment necessary for the cell culture includes a turntable, a pipette device, and a centrifuge tube handling device, and the culture operation required for the cell culture includes a medium exchange operation. And a passage operation can be included.

本発明の自動細胞培養装置の使用方法は、上記何れかの自動細胞培養装置の使用方法であって、一人の被検者についての前記培養操作を行った後、前記全ての密閉扉を閉じることにより、前記操作部のみの滅菌を行うことを特徴とする。   The method of using the automatic cell culture device of the present invention is a method of using any one of the above automatic cell culture devices, wherein after performing the culture operation for one subject, all the sealed doors are closed. Thus, only the operation unit is sterilized.

これにより、インキュベータ部に於いて複数の被検者の細胞培養を行うと共に、共通の操作部で複数の被検者についての個別の培養操作を交叉汚染を防止しつつ行うことが可能となる。   As a result, cell culture of a plurality of subjects can be performed in the incubator unit, and individual culture operations for the plurality of subjects can be performed in a common operation unit while preventing cross contamination.

また、本発明の自動細胞培養装置の使用方法は、上記何れかの自動細胞培養装置の使用方法であって、一人の被検者についての細胞培養が完了した後、前記インキュベータ部と前記操作部との間の密閉扉を開けると共に、当該密閉扉以外の密閉扉を閉じることにより、前記インキュベータ部及び前記操作部のみの滅菌を行うことを特徴とする。   Further, the method of using the automatic cell culture device of the present invention is a method of using any of the above automatic cell culture devices, and after the cell culture for one subject is completed, the incubator unit and the operation unit The sterilization of only the incubator part and the operation part is performed by opening the sealing door between and the sealing door other than the sealing door.

これにより、各インキュベータ部を異なる複数の被検者のために独立して使用することが可能となり、自動細胞培養装置の利用効率を高めることができる。   Thereby, it becomes possible to use each incubator part independently for several different subjects, and can improve the utilization efficiency of an automatic cell culture apparatus.

更に、本発明の自動細胞培養装置の使用方法は、上記何れかの自動細胞培養装置の使用方法であって、該自動細胞培養装置の保守時に、滅菌が必要な前記インキュベータ部、前記冷蔵保管部及び前記常温保管部と前記操作部との間の前記密閉扉を開いて、前記操作部と同時に前記インキュベータ部、前記冷蔵保管部及び前記常温保管部の滅菌を行うことを特徴とする。   Furthermore, the method of using the automatic cell culture device of the present invention is a method of using any of the above automatic cell culture devices, wherein the incubator unit and the refrigerated storage unit that require sterilization during maintenance of the automatic cell culture device And the said closed door between the said normal temperature storage part and the said operation part is opened, and the said incubator part, the said refrigerator storage part, and the said normal temperature storage part are sterilized simultaneously with the said operation part.

これにより、自動細胞培養装置の内部の滅菌が必要な部分の保守を簡単に行うことが可能となる。   As a result, it becomes possible to easily perform maintenance of a portion requiring sterilization inside the automatic cell culture apparatus.

ここで、上記の自動細胞培養装置の使用方法に於いては、前記入出庫部と前記操作部との間の前記密閉扉を更に開いて、前記入出庫部の滅菌を同時に行ってもよい。   Here, in the method of using the automatic cell culture apparatus, the sealing door between the loading / unloading unit and the operation unit may be further opened to sterilize the loading / unloading unit at the same time.

また、前記機器設置部と前記培養操作部との間に遮断装置を設けた自動細胞培養装置では、この遮断装置を閉じることにより、前記培養操作部のみの滅菌を行うことができる。   In addition, in an automatic cell culture device in which a shut-off device is provided between the device installation unit and the culture operation unit, only the culture operation unit can be sterilized by closing the shut-off device.

本発明の自動細胞培養装置ではオートクレーブ滅菌が採用されているため、ガス滅菌のように残留ガスの心配がなく、短時間で滅菌が完了する。しかも、オートクレーブ滅菌の採用により、病院等のような通常の医療機関で使用される滅菌装置と同様に高い滅菌効果が発揮される。   Since autoclave sterilization is employed in the automatic cell culture apparatus of the present invention, there is no concern about residual gas unlike gas sterilization, and sterilization is completed in a short time. In addition, by adopting autoclave sterilization, a high sterilization effect is exhibited as in a sterilization apparatus used in a normal medical institution such as a hospital.

また、密閉扉の選択的な開閉により、被検者ごと、培養操作ごとに必要な部分のみを滅菌することができるので、一台の装置で複数の被検者の細胞培養を行う装置でありながら、確実に交叉汚染を防止することができる。   In addition, by selectively opening and closing the sealing door, it is possible to sterilize only the necessary parts for each subject and for each culture operation, so it is a device that performs cell culture of multiple subjects with a single device However, cross contamination can be surely prevented.

本発明のオートクレーブ滅菌を利用した自動細胞培養装置及びその使用方法について、図面を参照しながら以下に説明するが、本発明は以下の説明に限定されるものではない。 Although the automatic cell culture apparatus using autoclave sterilization of the present invention and the method of using the same will be described below with reference to the drawings, the present invention is not limited to the following description.

図１は本発明の一実施形態に係る自動細胞培養装置の平面図であり、図１の装置をＰ−Ｐ線で破断して矢視右側上方から見た場合の斜視図を図２に示す。また、図３及び図４は、図１のＱ−Ｑ線矢視断面図を表している。本実施形態の自動細胞培養装置は、図１及び図２に示すように、操作部１１と、複数のインキュベータ部１４と、保管部としての冷蔵保管部１６及び常温保管部１７と、入出庫部１５と、蒸気供給部２２とを備えている。   FIG. 1 is a plan view of an automatic cell culture device according to an embodiment of the present invention, and FIG. 2 shows a perspective view when the device of FIG. . 3 and 4 show cross-sectional views taken along the line Q-Q in FIG. As shown in FIGS. 1 and 2, the automatic cell culture device of the present embodiment includes an operation unit 11, a plurality of incubator units 14, a refrigerated storage unit 16 and a room temperature storage unit 17 as storage units, and a loading / unloading unit. 15 and a steam supply unit 22.

操作部１１は被検者の細胞培養に必要な培養操作を行うためのスペースであり、図１に示すように、培養操作部１３とそれ以外の機器設置部１２とにより構成されている。培養操作部１３には、図１に示すように、ターンテーブル３１と、ピペット装置３２と、遠心管ハンドリング装置３３と、図示しない使用済み薬剤廃棄装置が設置されている。また、機器設置部１２には、人手に代わって実際の培養操作を行う操作ロボット１９と、遠心分離器２１とが設置されている。これらの操作ロボット１９、遠心分離器２１、ターンテーブル３１、ピペット装置３２、遠心管ハンドリング装置３３等は、図１に示すコントローラ２３の制御の下に動作している。   The operation unit 11 is a space for performing a culture operation necessary for the cell culture of the subject, and includes a culture operation unit 13 and other device installation unit 12 as shown in FIG. As shown in FIG. 1, the culture operation unit 13 is provided with a turntable 31, a pipette device 32, a centrifuge tube handling device 33, and a used medicine disposal device (not shown). In addition, the device installation unit 12 is provided with an operation robot 19 that performs an actual culture operation in place of a human hand, and a centrifuge 21. These operating robot 19, centrifuge 21, turntable 31, pipette device 32, centrifuge tube handling device 33, etc. operate under the control of the controller 23 shown in FIG.

インキュベータ部１４は細胞を培養するためのスペースであり、図３及び図４に示すように、インキュベータ部１４のそれぞれの内部には、細胞培養に使用される多数のシャーレ３４を保持する多段式のシャーレラック３５が配されている。また、図１に示すように、各インキュベータ部１４には、培養雰囲気を調整するための二酸化炭素ガスを供給する配管１４ａが接続されている。なお、図２では配管１４ａは２本のみ画かれているが、実際には全てのインキュベータ部１４に二酸化炭素ガスを供給する配管１４ａが接続されている。本実施形態では、一つのインキュベータ部１４内で培養される培養細胞は同一の被検者に由来するものに限定され、インキュベータ部１４ごとに異なる被検者の培養細胞を培養することができるようになっている。   The incubator unit 14 is a space for culturing cells. As shown in FIGS. 3 and 4, each incubator unit 14 has a multistage type holding a large number of petri dishes 34 used for cell culture. A petri dish 35 is arranged. As shown in FIG. 1, each incubator unit 14 is connected to a pipe 14 a that supplies carbon dioxide gas for adjusting the culture atmosphere. In FIG. 2, only two pipes 14 a are drawn, but actually, pipes 14 a that supply carbon dioxide gas to all incubator units 14 are connected. In the present embodiment, the cultured cells cultured in one incubator unit 14 are limited to those derived from the same subject, so that different cultured cells can be cultured for each incubator unit 14. It has become.

冷蔵保管部１６は、培養に必要な試薬類を冷蔵保存するために設けられており、この試薬類として、培地、培養細胞剥離用のトリプシン溶液等を例示することができる。冷蔵保管部１６には、既に滅菌済みの試薬類が収蔵されている。   The refrigerated storage unit 16 is provided for refrigerated storage of reagents necessary for culture, and examples of the reagents include a culture medium, a trypsin solution for detaching cultured cells, and the like. The refrigerated storage unit 16 stores already sterilized reagents.

常温保管部１７は、細胞培養に使用する培養器具類や常温保存が必要な薬剤を保管するために設けられており、この培養器具類として、培地交換に際して使用するピペットチップ、遠心管等を例示することができる。常温保管部１７には、既に滅菌済みの培養器具類が収蔵されている。さらに、常温保管部１７には常温保管が必要な薬品（例えばリン酸干渉バッファ（ＰＢＳ）や生理食塩水）などが収蔵されている。   The room temperature storage unit 17 is provided to store culture instruments used for cell culture and drugs that need to be stored at room temperature. Examples of the culture instruments include pipette tips and centrifuge tubes used for medium replacement. can do. The room temperature storage unit 17 stores already sterilized culture instruments. Further, the room temperature storage unit 17 stores chemicals that need to be stored at room temperature (for example, phosphate interference buffer (PBS) and physiological saline).

入出庫部１５は、培養すべき細胞、培養後の細胞、試薬類及び培養器具類の出し入れを行うために設けられている。さらに入出庫部は、培養する細胞の搬入口、培養後の搬出口としても利用しても良い。   The loading / unloading unit 15 is provided for loading / unloading cells to be cultured, cells after culture, reagents, and culture tools. Further, the loading / unloading unit may be used as a carrying-in port for cells to be cultured and a carrying-out port after culturing.

また、蒸気供給部２２はオートクレーブ滅菌を行うための蒸気を供給するものであり、例えば１２１℃の飽和蒸気（滅菌蒸気）を供給することができる。   The steam supply unit 22 supplies steam for performing autoclave sterilization, and can supply saturated steam (sterilized steam) at 121 ° C., for example.

本実施形態では、複数のインキュベータ部１４及び一つの入出庫部１５は、図２に示すように、上下に３段に設けられた複数の縦列を為して操作部１１に取り付けられている。入出庫部１５は、図２に示すように、上下３段の縦列の中央に位置している。冷蔵保管部１６及び常温保管部１７は上下２段の縦列を為し、上側に冷蔵保管部１６、下側に常温保管部１７が位置している。   In the present embodiment, the plurality of incubators 14 and the single storage / removal unit 15 are attached to the operation unit 11 in a plurality of columns provided in three stages up and down as shown in FIG. As shown in FIG. 2, the loading / unloading unit 15 is located at the center of three vertical columns. The refrigerated storage unit 16 and the room temperature storage unit 17 are arranged in two upper and lower columns, and the refrigeration storage unit 16 is positioned on the upper side and the room temperature storage unit 17 is positioned on the lower side.

また、本実施形態の自動細胞培養装置は、図２〜図４に示すように、操作部１１の機器設置部１２の上方にフィルタ部２０ａ、培養操作部１３の上方にフィルタ部２０ｂを備えている。フィルタ部２０ａ及び２０ｂは、ＨＥＰＡフィルタ（High Efficiency Particulate Air Filter）からなる。   Moreover, the automatic cell culture apparatus of this embodiment is equipped with the filter part 20a above the apparatus installation part 12 of the operation part 11, and the filter part 20b above the culture operation part 13, as shown in FIGS. Yes. The filter units 20a and 20b are composed of a HEPA filter (High Efficiency Particulate Air Filter).

本実施形態の装置では、操作部１１とインキュベータ部１４との間には自動開閉式の密閉扉１８ａ、操作部１１と入出庫部１５との間には自動開閉式の密閉扉１８ｂ、操作部１１と冷蔵保管部１６との間には自動開閉式の密閉扉１８ｃ、操作部１１と常温保管部１７との間には自動開閉式の密閉扉１８ｄがそれぞれ設けられている。また、図３及び図４に示すように、フィルタ部２０ａと機器設置部１２との間、及びフィルタ部２０ｂと培養操作部１３との間には、それぞれ自動開閉式の密閉扉３０ａ及び３０ｂが設けられている。更に、操作部１１の機器設置部１２及び培養操作部１３との間には、遮断装置としての密閉シャッター２８が設けられている。これらの密閉扉１８ａ〜ｄ,３０ａ及び３０ｂ、並びに密閉シャッター２８は、高い機密性と断熱性とを有する材料で形成されており、オートクレーブ滅菌の際には、操作部１１からの蒸気の漏れを防ぐとともに、操作部１１からの熱伝導を極力抑えることが可能となっている。   In the apparatus of the present embodiment, an automatic opening / closing type sealing door 18a is provided between the operation unit 11 and the incubator unit 14, and an automatic opening / closing type sealing door 18b is provided between the operation unit 11 and the loading / unloading unit 15. An automatic opening / closing type sealing door 18 c is provided between the refrigeration storage unit 16 and the refrigerator storage unit 16, and an automatic opening / closing type sealing door 18 d is provided between the operation unit 11 and the room temperature storage unit 17. Moreover, as shown in FIG.3 and FIG.4, between the filter part 20a and the apparatus installation part 12, and between the filter part 20b and the culture | cultivation operation part 13, the automatic opening-closing type sealing doors 30a and 30b are respectively provided. Is provided. Further, a hermetic shutter 28 as a shut-off device is provided between the device installation unit 12 and the culture operation unit 13 of the operation unit 11. These sealed doors 18a to 18d, 30a and 30b, and the sealed shutter 28 are formed of a material having high confidentiality and heat insulation properties, and prevent steam leakage from the operation unit 11 during autoclave sterilization. It is possible to prevent heat conduction from the operation unit 11 as much as possible.

従って、密閉扉１８ａ〜ｄが開いている状態では、インキュベータ部１４、入出庫部１５、冷蔵保管部１６及び常温保管部１７は操作部１１と空間的に連通し、蒸気供給部２２からの蒸気により操作部１１とともに滅菌されることとなるが、密閉扉１８ａ〜ｄを閉じたときには、インキュベータ部１４、入出庫部１５、冷蔵保管部１６及び常温保管部１７は操作部１１からほぼ完全に隔離されることとなり、滅菌蒸気の影響は及ばない。また、フィルタ部２０ａ及び２０ｂに設けられている密閉扉３０ａ及び３０ｂは、操作部１１の滅菌に際して閉じることにより、フィルタ部２０ａ及び２０ｂを滅菌蒸気から保護するために設けられている。更に、密閉シャッター２８は、これを閉じることにより、機器設置部１２又は培養操作部１３のみを滅菌することが可能となっている。本実施形態では、密閉扉１８ａ〜ｄ,３０ａ及び３０ｂの開閉、密閉シャッター２８の開閉もコントローラ２３の制御の下に行われる。   Therefore, in a state where the sealed doors 18a to 18d are open, the incubator unit 14, the loading / unloading unit 15, the refrigerated storage unit 16 and the room temperature storage unit 17 are in spatial communication with the operation unit 11 and steam from the steam supply unit 22 is obtained. However, when the closed doors 18a to 18d are closed, the incubator unit 14, the storage / removal unit 15, the refrigerated storage unit 16, and the room temperature storage unit 17 are almost completely separated from the operation unit 11. Therefore, the influence of sterilization steam is not affected. Moreover, the sealing doors 30a and 30b provided in the filter parts 20a and 20b are provided in order to protect the filter parts 20a and 20b from sterilization vapor | steam by closing when the operation part 11 is sterilized. Furthermore, the sealing shutter 28 can be sterilized only by the apparatus installation part 12 or the culture operation part 13 by closing this. In the present embodiment, opening and closing of the sealed doors 18 a to 18 d, 30 a and 30 b and opening and closing of the sealed shutter 28 are also performed under the control of the controller 23.

図３及び図４に示すように、本実施形態では操作部１１の機器設置部１２及び培養操作部１３には、給気用ファン２４及び排気用ファン２５により、ダウンフローの気流が供給されている。この気流は、給気用ファン２４によりプレフィルタ２４ａを介して吸引され、その大部分はフィルタ部２０ａで除塵された後に機器設置部１２に供給され、培養操作部１３の下方から排気用ファン２５によりプレフィルタ２５ａを介して外部に排気される。また、給気用ファン２４で吸引された気流の一部は、フィルタ部２０ｂで除塵された後に培養操作部１３に供給され、機器設置部１２の下方から排気用ファン２５を介して外部に排気される。フィルタ２５ａを通過した気流は、外部に廃棄することなく吸気用のフィルタ２４ａを介して後にＨＥＰＡフィルタ２０ａあるいは２０ｂを通すことによって、装置に戻す内部循環を行うことも可能である。機器設置部１２及び培養操作部１３に供給される気流の量は、シャッター２６及びダンパー２７により調節される。   As shown in FIGS. 3 and 4, in this embodiment, the apparatus installation unit 12 and the culture operation unit 13 of the operation unit 11 are supplied with the downflow airflow by the air supply fan 24 and the exhaust fan 25. Yes. The airflow is sucked by the air supply fan 24 through the prefilter 24 a, and most of the airflow is removed by the filter unit 20 a and then supplied to the device installation unit 12, and the exhaust fan 25 from below the culture operation unit 13. Thus, the air is exhausted outside through the prefilter 25a. Further, a part of the airflow sucked by the air supply fan 24 is supplied to the culture operation unit 13 after being dust-removed by the filter unit 20 b, and exhausted to the outside through the exhaust fan 25 from below the device installation unit 12. Is done. The airflow that has passed through the filter 25a can be internally circulated back to the apparatus by passing it through the HEPA filter 20a or 20b through the intake filter 24a without being discarded outside. The amount of airflow supplied to the device installation unit 12 and the culture operation unit 13 is adjusted by the shutter 26 and the damper 27.

本実施形態の装置では、細胞培養に必要な培養操作である培地交換操作と継代操作とを自動で行うことができる。本実施形態の自動細胞培養装置に於いては、培地交換操作は以下のようにして行われる。まず、図３に示すように、操作ロボット１９がインキュベータ部１４内のシャーレラック３５に載置されている細胞培養中のシャーレ３４を取り出し、図４に示すように培養操作部１３の方向に向きを変え、培養操作部１３のターンテーブル３１上にシャーレ３４を移動させる。次に、操作ロボット１９はシャーレ３４の蓋を開ける。続いて、ピペット装置３２がシャーレ内の培地を吸引して廃棄し、新たな培地をシャーレに供給する。次に、操作ロボット１９がシャーレ３４の蓋を閉じ、インキュベータ部１４内のシャーレラック３５の元の位置に戻す。このような一連の操作は、コントローラ２３の制御の下に行われる。   In the apparatus of the present embodiment, a medium exchange operation and a subculture operation, which are culture operations necessary for cell culture, can be automatically performed. In the automatic cell culture device of this embodiment, the medium exchange operation is performed as follows. First, as shown in FIG. 3, the operating robot 19 takes out the petri dish 34 in the cell culture placed on the petri dish rack 35 in the incubator unit 14, and faces the culture operating unit 13 as shown in FIG. And the petri dish 34 is moved onto the turntable 31 of the culture operation unit 13. Next, the operating robot 19 opens the lid of the petri dish 34. Subsequently, the pipette device 32 sucks and discards the medium in the petri dish, and supplies a new medium to the petri dish. Next, the operating robot 19 closes the lid of the petri dish 34 and returns it to the original position of the petri dish rack 35 in the incubator unit 14. Such a series of operations is performed under the control of the controller 23.

次に、本実施形態の自動細胞培養装置に於ける継代操作は、以下のようにして行われる。まず、前述と同様に操作ロボット１９がインキュベータ部１４内のシャーレラック３５に載置されている細胞培養中のシャーレ３４を取り出し、培養操作部１３のターンテーブル３１上にシャーレ３４を移動させ、シャーレ３４の蓋を開け、ピペット装置３２がシャーレ内の培養液を吸引して廃棄する。次に、ピペット装置３２は、培養細胞をシャーレから剥離するためのトリプシン溶液を滴下する。次に、操作ロボット１９はシャーレを傾けて回転することにより、トリプシン溶液をシャーレ３４の全体に行き渡らせる。細胞がシャーレから剥離したことが例えばＴＶカメラ（図示せず）等で確認されると、ピペット装置３２はシャーレ３４内の細胞を含んだ液を、遠心管ハンドリング装置３３に保持されている遠心管に移す。遠心管ハンドリング装置３３は遠心管にキャップをした後、これを遠心分離器２１にセットする。遠心分離器２１での遠心分離が終了すると、遠心管ハンドリング装置３３は遠心管からキャップを外す。次に、ピペット装置３２は遠心管の上澄み液を廃棄し、遠心管の底部に沈降している細胞を吸い出し、ターンテーブル３１上の複数の新たなシャーレに少量ずつ分配する。次に、操作ロボット１９はこれらのシャーレの蓋を閉じ、インキュベータ部１４内のシャーレラック３５に一つづつ入れ、その位置が次回の培地交換操作と継代操作のためにコントローラ２３に記憶される。   Next, the subculture operation in the automatic cell culture apparatus of this embodiment is performed as follows. First, in the same manner as described above, the operation robot 19 takes out the petri dish 34 in the cell culture placed on the petri dish rack 35 in the incubator unit 14, moves the petri dish 34 onto the turntable 31 of the culture operation unit 13, and moves the petri dish The lid 34 is opened, and the pipette device 32 sucks and discards the culture solution in the petri dish. Next, the pipette device 32 drops a trypsin solution for separating the cultured cells from the petri dish. Next, the operating robot 19 rotates the petri dish and rotates the trypsin solution throughout the petri dish 34. When it is confirmed by a TV camera (not shown) or the like that the cells have been detached from the petri dish, the pipette device 32 uses the centrifuge tube held in the centrifuge tube handling device 33 to remove the liquid containing the cells in the petri dish 34. Move to. The centrifuge tube handling device 33 caps the centrifuge tube and then sets it in the centrifuge 21. When centrifugation in the centrifuge 21 is completed, the centrifuge tube handling device 33 removes the cap from the centrifuge tube. Next, the pipette device 32 discards the supernatant of the centrifuge tube, sucks out the cells that have settled at the bottom of the centrifuge tube, and distributes them to a plurality of new dishes on the turntable 31 little by little. Next, the operation robot 19 closes the lids of these petri dishes, puts them one by one in the petri dish rack 35 in the incubator unit 14, and stores the positions in the controller 23 for the next medium exchange operation and subculture operation. .

本実施形態の自動細胞培養装置に於いては、オートクレーブ滅菌は、一人の被検者についての培養操作後、一人の被検者についての細胞培養が完了した後、自動細胞培養装置の保守時の他、培養細胞、試薬類、培養器具類等をこの装置内に搬入又はこの装置から搬出した時にも行われる。以下、本実施形態の自動細胞培養装置を用いたオートクレーブ滅菌の方法について説明する。   In the automatic cell culture apparatus of the present embodiment, autoclave sterilization is performed after a culture operation for one subject, after cell culture for one subject is completed, and after maintenance of the automatic cell culture apparatus. In addition, it is also performed when cultured cells, reagents, culture tools, etc. are carried into or out of this apparatus. Hereinafter, the autoclave sterilization method using the automatic cell culture apparatus of the present embodiment will be described.

図５〜図７は、図１のＰ−Ｐ線矢視断面図を表しており、これらの図に於いては、操作ロボット１９は省略してある。まず、一人の被検者について上記の培地交換操作、継代操作等の培養操作を行った後、別の被検者の培養操作を行う場合に行われるオートクレーブ滅菌について説明する。この場合、滅菌すべき場所は操作部１１のみなので、各インキュベータ部１４の密閉扉１８ａ、入出庫部１５の密閉扉１８ｂ、冷蔵保管部１６の密閉扉１８ｃ、常温保管部１７の密閉扉１８ｄ、並びにフィルタ部２０ａ及び２０ｂの密閉扉３０ａ及び３０ｂが閉じられる。次に、蒸気供給部２２から操作部１１に、例えば１２１℃の飽和蒸気が供給される（図６）。その際、密閉扉１８ａ〜ｄは前述のように高い機密性及び断熱性を有するので、各インキュベータ部１４内の培養中の細胞、冷蔵保管部１６及び常温保管部１７に収蔵されている試薬類、培養器具類に損傷を与えることはない。滅菌に十分な時間が経過した後、蒸気供給を止め、操作部１１が常温に戻れば、引き続いて別の被検者の細胞の培養操作を行うことが可能となる。   5 to 7 show cross-sectional views taken along the line P-P in FIG. 1, and the operation robot 19 is omitted in these drawings. First, autoclave sterilization, which is performed when one subject is subjected to the culture operation such as the medium replacement operation and the passage operation, and then another subject is subjected to the culture operation, will be described. In this case, since the operation part 11 is the only place to be sterilized, the sealed door 18a of each incubator part 14, the sealed door 18b of the loading / unloading part 15, the sealed door 18c of the refrigerated storage part 16, the sealed door 18d of the room temperature storage part 17, And the sealing doors 30a and 30b of the filter parts 20a and 20b are closed. Next, for example, saturated steam at 121 ° C. is supplied from the steam supply unit 22 to the operation unit 11 (FIG. 6). At that time, since the sealed doors 18a to 18d have high confidentiality and heat insulation properties as described above, the cells being cultured in each incubator unit 14, the reagents stored in the refrigerated storage unit 16 and the room temperature storage unit 17 are used. No damage to culture equipment. After a sufficient time for sterilization has elapsed, if the supply of steam is stopped and the operation unit 11 returns to room temperature, it becomes possible to continue the operation of culturing cells of another subject.

次に、一人の被検者についての細胞培養が完了した後に行われるオートクレーブ滅菌について説明する。この場合の滅菌は、培地交換操作、継代操作等の培養操作を繰り返して行うことにより、所定の程度までの培養が完了し、その培養細胞を取り出した後において行われるものである。この場合、滅菌すべき場所は操作部１１と特定のインキュベータ部１４のみなので、そのインキュベータ部１４の密閉扉１８ａを開けると共に、他の全ての密閉扉１８ａ〜ｄ、並びに密閉扉３０ａ及び３０ｂが閉じられる。次に、蒸気供給部２２から例えば１２１℃の飽和蒸気が供給される。その際、滅菌すべきインキュベータ部１４の密閉扉１８ａは開けられているので、滅菌蒸気により操作部１１及び当該インキュベータ部１４のみの滅菌が行われることとなる。滅菌に十分な時間が経過した後、蒸気供給を止め、当該インキュベータ部１４及び操作部１１が常温に戻れば、滅菌済みのインキュベータ部１４を用いて別人の細胞培養が開始されることとなる。   Next, autoclave sterilization performed after cell culture for one subject is completed will be described. The sterilization in this case is performed after completion of the culture to a predetermined degree by repeatedly performing culture operations such as medium exchange operation and subculture operation, and taking out the cultured cells. In this case, since only the operation unit 11 and the specific incubator unit 14 are to be sterilized, the sealed door 18a of the incubator unit 14 is opened and all the other sealed doors 18a to 18d and the sealed doors 30a and 30b are closed. It is done. Next, for example, saturated steam at 121 ° C. is supplied from the steam supply unit 22. At that time, since the sealed door 18a of the incubator unit 14 to be sterilized is opened, only the operation unit 11 and the incubator unit 14 are sterilized by sterilization steam. After a sufficient time for sterilization has elapsed, when the steam supply is stopped and the incubator unit 14 and the operation unit 11 return to room temperature, another person's cell culture is started using the sterilized incubator unit 14.

更に、オートクレーブ滅菌は、自動細胞培養装置の保守時にも行われる。この場合の滅菌は、冷蔵保管部１６の密閉扉１８ｃ及び常温保管部１７の密閉扉１８ｄの何れか一方又は両方と、滅菌が必要なインキュベータ部１４の密閉扉１８ａを開き、他の全ての密閉扉１８ａ〜ｂ、並びに密閉扉３０ａ及び３０ｂが閉じられる。この状態で、蒸気供給部２２から例えば１２１℃の飽和蒸気が供給される。これにより、操作部１１と、冷蔵保管部１６、常温保管部１７及び滅菌すべきインキュベータ部１４とが同時に滅菌されることとなる。なお、図７は、インキュベータ部１４及び常温保管部１７の滅菌は行わず、冷蔵保管部１６及び操作部１１のみの滅菌を行う場合を表している。   Furthermore, autoclave sterilization is also performed during maintenance of the automatic cell culture apparatus. In this case, sterilization is performed by opening one or both of the sealed door 18c of the refrigerated storage unit 16 and the sealed door 18d of the room temperature storage unit 17 and the sealed door 18a of the incubator unit 14 that requires sterilization. The doors 18a to 18b and the sealed doors 30a and 30b are closed. In this state, for example, saturated steam at 121 ° C. is supplied from the steam supply unit 22. Thereby, the operation part 11, the refrigerator storage part 16, the normal temperature storage part 17, and the incubator part 14 which should be sterilized will be sterilized simultaneously. FIG. 7 shows a case where only the refrigerated storage unit 16 and the operation unit 11 are sterilized without sterilization of the incubator unit 14 and the room temperature storage unit 17.

また、本実施形態の自動細胞培養装置に於いては、入出庫部１５のオートクレーブ滅菌も適宜行われる。入出庫部１５の滅菌は、入出庫部１５の密閉扉１８ｂを開けておくことにより行うことができ、培養細胞、試薬類、培養器具類等をこの装置内に搬入又はこの装置から搬出した時に随時行うことができ、上述の培養操作後の操作部１１の滅菌、一人の被検者の細胞培養完了後に於ける操作部１１とインキュベータ部１４の滅菌、自動細胞培養装置の保守時の滅菌等と同時に行うことができる。   Moreover, in the automatic cell culture apparatus of this embodiment, the autoclave sterilization of the loading / unloading part 15 is also performed suitably. The sterilization of the loading / unloading unit 15 can be performed by opening the sealed door 18b of the loading / unloading unit 15, and when cultured cells, reagents, culture tools, etc. are carried into or out of this device. Sterilization of the operation unit 11 after the above-described culture operation, sterilization of the operation unit 11 and the incubator unit 14 after completion of cell culture of one subject, sterilization during maintenance of the automatic cell culture apparatus, etc. Can be done at the same time.

加えて、本実施形態の自動細胞培養装置では、機器設置部１２と培養操作部１３との間に密閉シャッター２８が設けられているので、蒸気供給部２２からの滅菌蒸気を培養操作部１３のみに導入するようにすれば、密閉シャッター２８を閉じることにより、培養操作部１３のみの滅菌を行うことができる。また、蒸気供給部２２からの滅菌蒸気を機器設置部１２のみに導入するようにすれば、密閉シャッター２８を閉じることにより、機器設置部１２のみの滅菌を行うこともできる。   In addition, in the automatic cell culture device of the present embodiment, since the sealing shutter 28 is provided between the device installation unit 12 and the culture operation unit 13, only the culture operation unit 13 is supplied with sterilized steam from the steam supply unit 22. In this case, only the culture operation unit 13 can be sterilized by closing the sealing shutter 28. Further, if the sterilization steam from the steam supply unit 22 is introduced only into the device installation unit 12, only the device installation unit 12 can be sterilized by closing the sealing shutter 28.

更に、本実施形態の自動細胞培養装置は、その内部全体の滅菌を行うこともできる。即ち、密閉扉３０ａ及び３０ｂを閉じるとともに全ての密閉扉１８ａ〜ｄを開けることにより、操作部１１の機器設置部１２及び培養操作部１３、インキュベータ部１４、入出庫部１５、冷蔵保管部１６並びに常温保管部１７を同時に滅菌することができる。   Furthermore, the automatic cell culture apparatus of this embodiment can also sterilize the whole inside. That is, by closing the sealing doors 30a and 30b and opening all the sealing doors 18a to 18d, the equipment installation unit 12 and the culture operation unit 13, the incubator unit 14, the loading / unloading unit 15, the refrigerated storage unit 16 of the operation unit 11 and The room temperature storage unit 17 can be sterilized at the same time.

本発明のオートクレーブ滅菌を利用した自動細胞培養装置及びその使用方法によれば、細胞の培養操作を自動で行うことができるので、再生医療の分野で利用することができる。   According to the automatic cell culturing apparatus using autoclave sterilization and the method of using the same of the present invention, the cell culturing operation can be performed automatically, so that it can be used in the field of regenerative medicine.

本発明の一実施形態に係る自動細胞培養装置の平面図である。It is a top view of the automatic cell culture device concerning one embodiment of the present invention. 図１の装置をＰ−Ｐ線で破断して矢視右側上方から見た場合の斜視図である。It is a perspective view at the time of fracture | rupturing the apparatus of FIG. 1 by PP line and seeing from the arrow right upper side. 図１のＱ−Ｑ線矢視断面図である。It is the QQ arrow directional cross-sectional view of FIG. 図１のＱ−Ｑ線矢視断面図である。It is the QQ arrow directional cross-sectional view of FIG. 図１のＰ−Ｐ線矢視断面図である。FIG. 2 is a cross-sectional view taken along line P-P in FIG. 1. 図５に於いて、蒸気供給部２２から操作部１１に滅菌蒸気を供給した場合を表している。In FIG. 5, the case where sterilization vapor | steam is supplied to the operation part 11 from the vapor | steam supply part 22 is represented. 図１のＰ−Ｐ線矢視断面図であり、冷蔵保管部１６と操作部１１とを滅菌する場合を表している。FIG. 2 is a cross-sectional view taken along the line P-P in FIG. 1 and illustrates a case where the refrigerated storage unit 16 and the operation unit 11 are sterilized.

符号の説明Explanation of symbols

１１ 操作部
１２ 機器設置部
１３ 培養操作部
１４ インキュベータ部
１５ 入出庫部
１６ 冷蔵保管部
１７ 常温保管部
１８ａ〜ｄ 密閉扉
３０ａ，３０ｂ 密閉扉
１９ 操作ロボット
２０ａ，２０ｂ フィルタ部
２１ 遠心分離器
２２ 蒸気供給部
２３ コントローラ
２４ 給気用ファン
２４ａ，２５ａ プレフィルタ
２５ 排気用ファン
２６ シャッター
２７ ダンパー
２８ 密閉シャッター
３１ ターンテーブル
３２ ピペット装置
３３ 遠心管ハンドリング装置
３４ シャーレ
３５ シャーレラック
11 Operation unit
12 Equipment installation section
13 Culture operation part
14 Incubator
15 Entry / Exit Department
16 Refrigerated storage
17 Room temperature storage part 18a-d Sealed door 30a, 30b Sealed door
19 Operation robot 20a, 20b Filter part
21 Centrifuge
22 Steam supply section
23 Controller
24 Air supply fan 24a, 25a Pre-filter
25 Exhaust fan
26 Shutter
27 Damper
28 Sealed shutter
31 Turntable
32 pipette device
33 Centrifuge tube handling equipment
34 Petri dish
35 petri dish rack

Claims (12)

被検者の細胞培養に必要な培養操作を行うための操作部と、細胞を培養するためのインキュベータ部と、培養に必要な試薬類や培養器具類を保管する保管部と、オートクレーブ滅菌を行うための蒸気供給部とを備え、
前記インキュベータ部及び前記保管部は、それぞれ前記操作部に連通すると共に、前記操作部との間にそれぞれ密閉扉を備えており、該密閉扉を選択して開閉することにより、前記操作部と、該操作部に通ずる前記インキュベータ部及び前記保管部の何れかとに前記蒸気供給部からの滅菌蒸気が供給されることを特徴とする自動細胞培養装置。 An operation unit for performing culture operations necessary for cell culture of a subject, an incubator unit for culturing cells, a storage unit for storing reagents and culture tools necessary for culture, and autoclave sterilization A steam supply section for
The incubator unit and the storage unit communicate with the operation unit, respectively, and each has an airtight door between the operation unit, and by selecting and opening and closing the airtight door, the operation unit, The automatic cell culture apparatus, wherein sterilization vapor from the vapor supply unit is supplied to either the incubator unit or the storage unit that communicates with the operation unit. 試薬及び培養器具類の出し入れを行うための入出庫部を更に備え、
前記入出庫部は前記操作部に連通すると共に、前記操作部との間に密閉扉を備えており、前記密閉扉を開閉することにより前記蒸気供給部からの滅菌蒸気が供給されることを特徴とする請求項１記載の自動細胞培養装置。 It further includes a loading / unloading unit for loading and unloading reagents and culture equipment,
The entry / exit unit communicates with the operation unit and includes a sealed door between the operation unit and the sterilization steam from the steam supply unit is supplied by opening and closing the seal door. The automatic cell culture apparatus according to claim 1 . 前記操作部に供給される気流の除塵及び除菌を行うフィルタ部と、該フィルタ部及び前記操作部の間に設けられた密閉扉とを更に備え、オートクレーブ滅菌に際して前記フィルタ部を前記操作部から遮断することを特徴とする請求項１又は２記載の自動細胞培養装置。   A filter unit that performs dust removal and sterilization of the airflow supplied to the operation unit, and a sealed door provided between the filter unit and the operation unit, the filter unit being removed from the operation unit during autoclave sterilization The automatic cell culture device according to claim 1 or 2, wherein the automatic cell culture device is blocked. 前記操作部は、操作ロボットと、遠心分離器と、細胞培養に必要な培養操作機器とを備えている請求項１乃至３の何れかに記載の自動細胞培養装置。 The operation unit includes an operation robot, a centrifugal separator, the automatic cell cultivation apparatus according to any one of claims 1 to 3 and a culture operation equipment required for cell culture. 前記操作部は、前記操作ロボット及び前記遠心分離器を配置した機器設置部と、前記培養操作機器を配置した培養操作部とを備え、前記機器設置部と前記培養操作部との間に遮断装置が設けられている請求項４記載の自動細胞培養装置。   The operation unit includes a device installation unit in which the operation robot and the centrifuge are arranged, and a culture operation unit in which the culture operation device is arranged, and a blocking device is provided between the device installation unit and the culture operation unit. The automatic cell culture apparatus according to claim 4, wherein: 前記培養操作機器には、ターンテーブルと、ピペット装置と、遠心管ハンドリング装置とが含まれる請求項４又は５記載の自動細胞培養装置。   The automatic cell culture device according to claim 4 or 5, wherein the culture operation device includes a turntable, a pipette device, and a centrifuge tube handling device. 前記細胞培養に必要な前記培養操作には、培地交換操作と継代操作とが含まれる請求項１又は２記載の自動細胞培養装置。   The automatic cell culture apparatus according to claim 1 or 2, wherein the culture operation necessary for the cell culture includes a medium exchange operation and a passage operation. 請求項１乃至７の何れかの自動細胞培養装置の使用方法であって、一人の被検者についての前記培養操作を行った後、前記全ての密閉扉を閉じることにより、前記操作部のみの滅菌を行う自動細胞培養装置の使用方法。   The method for using the automatic cell culture apparatus according to any one of claims 1 to 7, wherein after performing the culture operation for one subject, only the operation unit is closed by closing all the sealed doors. How to use an automatic cell culture device for sterilization 請求項１乃至７の何れかの自動細胞培養装置の使用方法であって、一人の被検者についての細胞培養が完了した後、前記インキュベータ部と前記操作部との間の密閉扉を開けると共に、当該密閉扉以外の密閉扉を閉じることにより、前記インキュベータ部及び前記操作部のみの滅菌を行うことを特徴とする自動細胞培養装置の使用方法。   The method for using the automatic cell culture device according to any one of claims 1 to 7, wherein after the cell culture for one subject is completed, the sealed door between the incubator unit and the operation unit is opened. The method for using an automatic cell culture apparatus, wherein only the incubator part and the operation part are sterilized by closing a sealing door other than the sealing door. 請求項１乃至の７何れかの自動細胞培養装置の使用方法であって、該自動細胞培養装置の保守時に、滅菌が必要な前記インキュベータ部、前記保管部と前記操作部との間の前記密閉扉を開いて、前記操作部と同時に前記インキュベータ部、前記保管部の滅菌を行うことを特徴とする自動細胞培養装置の使用方法。   The method for using the automatic cell culture device according to any one of claims 1 to 7, wherein the sealing between the incubator unit, the storage unit, and the operation unit that requires sterilization during maintenance of the automatic cell culture device. A method for using an automatic cell culture device, wherein the door is opened and the incubator unit and the storage unit are sterilized simultaneously with the operation unit. 前記入出庫部と前記操作部との間の前記密閉扉を更に開いて、前記入出庫部の滅菌を同時に行うことを特徴とする請求項８乃至１０の何れかに記載の自動細胞培養装置の使用方法。   The automatic cell culture apparatus according to any one of claims 8 to 10, wherein the closed door between the storage / exit section and the operation section is further opened to sterilize the storage / exit section at the same time. how to use. 請求項５記載の自動細胞培養装置の使用方法であって、前記機器設置部と前記培養操作部との間の遮断装置を閉じることにより、前記培養操作部のみの滅菌を行うことを特徴とする自動細胞培養装置の使用方法。   6. The method for using an automatic cell culture device according to claim 5, wherein only the culture operation unit is sterilized by closing a shut-off device between the device installation unit and the culture operation unit. How to use automatic cell culture equipment.
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